Single-step purification of immunoglobulin M on C1q-Sepharose.
A rapid and simple affinity chromatography method for purifying IgM from myeloma serum and ascites fluid is described. Complement protein C1q is coupled to Sepharose with an efficiency of 35%, giving 1.7 mg of C1q bound/ml of gel. This C1q-Sepharose selectively binds IgM from crude samples at 5 degrees C, with a capacity of 0.4 mg of IgM/ml of gel. The bound IgM may be eluted simply and isocratically by bringing the gel to room temperature for 2 h, or by washing with buffer containing 0.5 M KI. The eluted IgM is highly pure by SDS-PAGE and double immunodiffusion analysis, although IgG may be a potential contaminant. The C1q-Sepharose is stable for at least 18 months.
Nethery
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A
Clinical Immunology Research Centre, University of Sydney, NSW, Australia.
Raison
R L
RL
Easterbrook-Smith
S B
SB
D002846:Chromatography, Affinity; D015922:Complement C1q / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007075:Immunoglobulin M / Q000302:isolation & purification*; D012685:Sepharose
D007075:Immunoglobulin M; D015922:Complement C1q; D012685:Sepharose